EIF2B bodies and their role in the integrated stress response

Abstract

Eukaryotic initiation factor 2 (eIF2) is a G protein comprised of 3 subunits (α, β and γ) that is critical for translation. It is tightly regulated in the integrated stress response (ISR) via the phosphorylation of its α subunit following the induction of cellular stress. In its phosphorylated form eIF2α inhibits the guanine nucleotide exchange factor (GEF) eukaryotic initiation factor 2B (eIF2B), resulting in the attenuation of global protein synthesis. eIF2B is a multisubunit protein comprised of regulatory and catalytic subunits. The catalytic subunits are responsible for the GEF activity whereas the regulatory subunits mediate inhibition by phosphorylated eIF2α. Through studying the localisation of eIF2B subunits, cytoplasmic eIF2B bodies were identified in mammalian cells. A relationship between body size and the eIF2B subunits localising to them exists; larger bodies contain all subunits and smaller bodies contain predominantly catalytic subunits. eIF2 localises to eIF2B bodies and moves through these bodies in a manner that correlates with eIF2B GEF activity. Upon the induction of cellular stress phosphorylated eIF2α localises predominately to larger eIF2B bodies which contain regulatory subunits and a decrease in the movement of eIF2 through these bodies is observed. Interestingly, drugs that inhibit the ISR can rescue the movement of eIF2 through these eIF2B bodies, in a manner that correlates to cellular levels of phosphorylated eIF2α. In contrast, smaller eIF2B bodies, which contain predominately catalytic subunits, show increased movement of eIF2 during cellular stress. This increase in movement is accompanied by an increase in the localisation of eIF2Bδ to these bodies, suggesting the formation of a novel eIF2B subcomplex. This response is mimicked by ISR-inhibiting drugs, providing insight into their potential mechanisms of action. This study provides the first evidence that the composition and function of mammalian eIF2B bodies is regulated by the ISR and the drugs that control it.